Abstract：
Adrenomedullin (ADM) has beneficial effects on Leydig cells under pathological conditions, including lipopolysaccharide (LPS)-induced orchitis. Our previous studies demonstrated that ADM exerts a restorative effect on steroidogenesis in LPS-treated primary rat Leydig cells by attenuating oxidative stress, inflammation and apoptosis. In this study, we aim to investigate whether ADM inhibits Leydig cell dysfunction by rescuing steroidogenic enzymes in vivo. Rats were administered with LPS and injected with Ad-ADM, an adeno-associated virus vector that expressed ADM. Then, rat testes were collected for 3β-hydroxysteroid dehydrogenase (3β-HSD) immunofluorescence staining. Steroidogenic enzymes or steroidogenic regulatory factors or protein, including steroidogenic factor-1 (SF-1), liver receptor homologue-1 (LRH1), Nur77, steroidogenic acute regulatory protein (StAR), cytochrome P450 cholesterol side chain cleavage enzyme (P450scc), 3β-HSD, cytochrome P450 17α-hydroxylase/17, 20 lyase (CYP17) and 17β-hydroxysteroid dehydrogenase (17β-HSD), were detected via gene expression profiling and western blot analysis. Plasma testosterone concentrations were measured. Results showed that ADM may inhibit Leydig cell dysfunction by rescuing steroidogenic enzymes and steroidogenic regulatory factors in vivo. The reduction in the number of Leydig cells after LPS exposure was reversed by ADM. ADM rescued the gene or protein levels of SF-1, LRH1, Nur77, StAR, P450scc, 3β-HSD, CYP17 and 17β-HSD and plasma testosterone concentrations. To summarize ADM could rescue some important steroidogenic enzymes, steroidogenic regulatory factors and testosterone production in Leydig cells in vivo.
摘要：
肾上腺髓质素（ADM）在病理条件下对睾丸间质细胞具有有益作用，包括脂多糖(LPS)诱导的睾丸炎。我们先前的研究表明，ADM通过减弱氧化应激对LPS处理的原代大鼠睾丸间质细胞的类固醇生成产生恢复作用，炎症和细胞凋亡。在这项研究中,我们旨在研究ADM是否通过在体内挽救类固醇生成酶来抑制Leydig细胞功能障碍。给大鼠施用LPS并注射Ad-ADM,表达ADM的腺相关病毒载体。然后，收集大鼠睾丸进行3β-羟基类固醇脱氢酶（3β-HSD）免疫荧光染色。类固醇生成酶或类固醇生成调节因子或蛋白质，包括类固醇生成因子-1（SF-1），肝受体同源物-1（LRH1），Nur77，类固醇生成急性调节蛋白（StAR），细胞色素P450胆固醇侧链裂解酶（P450scc），3β-HSD,细胞色素P45017α-羟化酶/17，20裂解酶（CYP17）和17β-羟基类固醇脱氢酶（17β-HSD），通过基因表达谱分析和蛋白质印迹分析进行检测。测量血浆睾酮浓度。结果表明，ADM可能通过挽救体内类固醇生成酶和类固醇生成调节因子来抑制Leydig细胞功能障碍。LPS暴露后Leydig细胞数量的减少被ADM逆转。ADM拯救了SF-1，LRH1，Nur77，StAR，P450scc,3β-HSD,CYP17和17β-HSD和血浆睾酮浓度。总结ADM可以拯救一些重要的类固醇生成酶，体内Leydig细胞中类固醇生成调节因子和睾酮的产生。
